Growth and Transcriptomics Analysis ofMichelia macclurei Dandy Plantlets with Different LED Quality Treatments

Michelia macclurei Dandy is a significant tree species that has extensive cultivation for forestry and horticulture purposes in southern China,owing to its economic and practical importance.Light quality influences plantlet growth and development during tissue culture.However,the growth characteristic and molecular regulation of M.macclurei under different light quality conditions are not well understood yet.In this study,we investigated the morphological,chlorophyll content,and transcriptomic responses of M.macclurei plantlets under different light-emitting diode(LED)qualities,including white,blue,and red light.The results showed that blue light significantly increased plant height(21.29%)and leaf number(18.65%),while red light decreased plant height and leaf number by 7.53%and 16.49%,respectively.In addition,the plantlets’chlorophyll content and etiolation rate were significantly reduced by blue and red light quality compared to white light.Compared to white light,blue light had a negative effect,leading to decreased rooting rate(64.28%),root number(72.72%),and root length(75.86%).Conversely,red light had a positive effect,resulting in increased rooting rate(24.99%),root number(109.58%),and root length(72.72%).Transcriptome analysis identified 54 differentially expressed genes(DEGs)in three groups that consisted of blue light vs.white light(BL-vs-WL),red light vs.white light(RL-vs-WL),and red light vs.blue light(RL-vs-BL).Specifically,21,7,and 41 DEGs were identified in the three groups,respectively.The DEGs found in the RL-vs-WL and BL-vs-WL groups were involved in plant hormone signaling,nitrogen metabolism,and phenylpropanoid biosynthesis pathways,which suggests that M.macclurei plantlets adapt to the changes of light quality via modulating gene expression.Overall,our study provides valuable insights for understanding the molecular and morphological responses of M.macclurei plantlets under different light qualities.

Evaluation of Different Substrates Compositions for Acclimatization of Tissue Culture Taro Plantlets in a Propagator

Taro is cultivated in most Regions of Cameroon and it is affected by taro leaf blight disease since 2010 which has decreased its production. Lack of disease-free planting materials has been a main problem to farmers. This study was carried out at International Institute of Tropical Agriculture (IITA) Yaounde and Institute of Agricultural Research for Development (IRAD) Bambui to assess different substrates for acclimatization of tissue culture taro plantlets in apropagator. No information is available on acclimatization of Cameroonian taro plantlets in different substrates. Taro plantlets from tissue culture were acclimatised in a propagator for six weeks under different substrates, the first substrate consisted of sterile three parts of soil and one part of river sand mixed together (3:1), the second substrate consisted of sterile two parts of soil and two parts of river sand mixed together (2:2), the third substrate consisted of sterile two parts of soil, one part of rice husk and one part of river sand mixed together (2:1:1) and the fourth substrate consisted of sterile one part of soil and three parts of river sand mixed together (1:3). After acclimatisation of the different taroplantlets (Dark green petiole with small leaves (L1), Red petiole with small leaves (L2), Light green petiole with large leaves (L3) and Light green petiole with small leaves (L4) in these four substrates, it was observed that the best growth rate of plant was recorded on substrate sand + soil (1:3). The other substrates showed moderate growth of plants. Substrate sand + soil (1:3) can be recommended for acclimatization of Cameroonian taro plantlets.

碳离子束辐照对马铃薯组培苗生长及生理特性的影响

为研究高能碳离子束对马铃薯组培苗茎切段的影响,以马铃薯品种‘新大坪’的组培苗茎切段为研究材料,设置4个辐照剂量梯度对其进行辐照处理,测定植株致死率、生根率、侧芽形成率、形态变异率以及叶片生理指标,确定‘新大坪’组培苗茎切段的半致死辐照剂量。结果表明:辐照剂量与致死率之间的关系呈正相关性;组培苗茎切段的再生植株表型变化主要为叶片不对称、叶缘不规则、茎变异伸长、叶片丛生和叶片粘连等;随辐照剂量升高,超氧化物歧化酶、过氧化物酶和过氧化氢酶活性呈先升后降的趋势;丙二醛含量则呈先升后降,随后又升高的现象。利用线性回归方程计算‘新大坪’的半致死辐照剂量为20.37 Gy。

珙桐体细胞胚胎发生技术研究

研究探讨基本培养基类型对珙桐愈伤组织诱导的影响、植物生长调节剂对胚性愈伤组织诱导及体胚发育的影响,初步建立以未成熟合子胚为外植体,通过体细胞胚胎发生途径再生珙桐植株的培养方案。结果表明,在含有3%蔗糖、0.3%凝胶、800 mg/L水解酪蛋白、400 mg/L L-谷氨酰胺的MS、1/2MS和WPM基本培养基上均可诱导出愈伤组织,以MS+2.0 mg/L 2,4-D+0.5 mg/L 6-BA诱导率最高,可达94.0%;胚性愈伤组织在MS+0.5 mg/L NAA+1.0 mg/L 6-BA的培养基上诱导效果较好;在MS+0.5 mg/L NAA+1.0 mg/L 6-BA培养基上,体胚诱导率可达22.9%;成熟体胚在1/2MS+0.5 mg/L 6-BA+0.25 mg/L IBA的萌发培养基上能正常萌发,其后在添加1 g/L活性炭的萌发培养基上转化成完整小植株。组织细胞学观察表明,珙桐体胚起源于胚性愈伤组织,经历了球形期、心形期、鱼雷期和子叶期发育阶段,这与自然条件下的合子胚形成过程相似。

In vitro callus induction and plantlet regeneration of Achyranthes aspera L.,a high value medicinal plant

Objective:To study callus induction from different explants(internode,leaf,root)and in vitro plantlets propagation from medicinally important plant Achyranthes aspera L.Methods:Sterilized explants were prepared by uning 0.1%HgCl_2 and 0.5%Bavistin and callus was obtained when cultured onto Murashige Skoog's(MS)medium by using different concentrations and combination of 2,4-D.NAA.BAP,IAA,IBA with 3%sucrose and 0.8%agar.Induced callus was immediately transferred to MS medium containing at different concentrations of phytohormones for shootlets and rootlets induction respectively.Results:Sterilization treatment of 0.1%HgCl_2.for 2-3 min and Bavistin 0.5%for 10-12 min showed the highest percentage of asepsis and survival rate.Maximum induction of callus was obtained from a combination of 2.0 mg/L 2,4-D and 0.5 mg/L NAA from leaf.Highest shootlets number(4.83±0.l7)and length(3.8±0.16)cm were observed on full strength MS medium when fortified with BAP 4.0 mg/L and KIN 0.5 mg/L.Concerted efforts of BAP 10 mg/L and NAA 0.5 mg/L on full strength MS medium showed highest leaf number(6.77±0.94).In vitro raised shoots were allowed to root on different strengths of MS medium fortified with IAA and IBA at different concentrations.Experimentally,3.0 mg/L IBA was enabled to induce maximum rootlets number(10.0±9.82)on full strength MS medium.Afterwards,regenerated shoots with well developed roots were successfully subjected to hardening process and were acclimatized.The survived plantlets showed 66.67%survival frequency without any morphological abnormality.Conclusions:The results demonstrated that different explants were good source of callus induction,morphology analysis as well as indirect plantlets regeneration.

Effects of different LEDs light spectrum on the growth,leaf anatomy,and chloroplast ultrastructure of potato plantlets in vitro and minituber production after transplanting in the greenhouse

Light spectrum plays an important role in regulating the growth and development of in vitro cultured potato(Solanum tuberosum L.) plantlets. The status of potato plantlets at the end of in vitro stage influences the minituber production after transplanting. With 100 μmol m^-2s^-1 total photosynthetic photon flux density(PPFD), a light spectrum study of 100% red light emitting diodes(LEDs) light spectrum(RR), 100% blue LEDs light spectrum(BB), 65% red+35% blue LEDs light spectrum(RB), and 45% red+35% blue+20% green LEDs light spectrum(RBG) providing illumination at the in vitro cultured stage of potato plantlets for 4 weeks using fluorescent lamp as control(CK) was performed to investigate the effects of LEDs light spectrum on the growth, leaf anatomy, and chloroplast ultrastructure of potato plantlets in vitro as well as the minituber yield after 2 months transplanting in the greenhouse. Compared to CK, RB and RBG promoted the growth of potato plantlets in vitro with increased stem diameter, plantlet fresh weight, plantlet dry weight, and health index. Furthermore, BB induced the greatest stem diameter as well as the highest health index in potato plantlets in vitro. Root activity, soluble protein, and free amino acid were also significantly enhanced by BB, whereas carbohydrates were improved by RR. In addition, thickness of leaf, palisade parenchyma and spongy parenchyma was significantly increased by BB and RBG. Chloroplasts under BB and RBG showed well-developed grana thylakoid and stroma thylakoid. Unexpectedly, distinct upper epidermis with greatest thickness was induced and palisade parenchyma and spongy parenchyma were arranged neatly in RR. After transplanting in the greenhouse for 2 months, potato plantlets in vitro from BB, RB, and RBG produced high percentage of large size tuber. BB improved fresh and dry weights of the biggest tuber but decreased tuber number per plantlet. In addition, RBG increased tuber number as well as tuber fresh and dry weight slightly. Our results suggested monochromatic blue LEDs as well as combined red, blue or/and green LEDs light spectrum were superior to fluorescent lamp spectrum in micro-propagation of potato plantlets. Therefore, the application of RBG was suitable;BB and RB could be used as alternatives.

Effect of mini-cutting size on adventitious rooting and morphophysiological quality of Ilex paraguariensis plantlets

As the energizing and therapeutic effects of infusions of mate(Ilex paraguariensis A.St.-Hil)leaves are increasingly recognized,rising global consumption is also increasing the demand for plantlets with high genetic and morphological qualities.Mini-cuttings are an alternative for clonal plantlets;however,more details are needed on factors that affect the success of this technique.Here,toward maximizing production,we evaluated adventitious rooting competence of mini-cuttings of different sizes and the morphophysiological quality of the resultant plantlets.Small(one bud;length up to 2.5 cm),medium(multiple buds;length 2.6–5.0 cm),and large(multiple buds;length 5.1–10.0 cm)mini-cuttings were planted in a mixture of commercial substrate,vermiculite,and coarse sand(1:1:1 v:v:v),grown in a humidity chamber,then evaluated after 45,60,75,and 90 days for survival,percentage of callus and shoot formation,number and length of shoots,percentage of rooted cuttings,and number and length of roots.Any rooted mini-cuttings were then cultivated in commercial substrate and subsurface soil(2:1 v:v)and evaluated after 30,60,90 and 120 days of cultivation for morphophysiology of the plantlets,percentage of survival,number of leaves,shoot height,stem diameter,and ratio of shoot height to stem diameter.After 120 days,the plantlets were evaluated for the ratio of shoot and root dry mass,total length,surface area,total volume,and mean diameter of roots and Dickson quality index.Adventitious rooting of the mini-cuttings and the morphophysiological quality of the plantlets were not influenced by the size of the vegetative propagule.Therefore,single-bud mini-cuttings have similar adventitious rooting competence,and result in plantlets with adequate morphophysiological quality,based upon leaf development,shoot height,stem diameter and length,and surface area,volume and diameter of roots.Single-bud mini-cuttings up to 2.5 cm long can thus be used to maximize production of mate plantlets by mini-cuttings.

Synthetic Seed Preparation, Germination and Plantlet Regeneration of Litchi (Litchi chinensis Sonn.)

Litchi chinensis sonn.) ranks second after mango amongst the most important fruit crops cultivated worldwide. Litchi is a very valuable crop throughout the world because it is a table fruit and wines are also produced from it. The existing cultivars are highly polyploidy and heterozygous in nature. It is propagated through air layering and marcottage methods and storability is very low. Synthetic seeds can be stored for a long time and its genetic constitution could remain the same. For germplasm maintenance and clonal propagation, synthetic seeds can be used. Somatic embryogenesis has been reported from anther or embryogenic suspension culture in various species of litchi. Regeneration via organogenesis and somatic embryogenesis from zygotic embryos has also been reported in certain species. Developing a methodology for getting somatic embryogenesis with a high frequency from zygotic embryos which is available once in a year, would be particularly useful for genetic improvement of litchi. Cotyledonary stage somatic embryos developed from zygotic embryos were encapsulated in 2% alginate gel. The encapsulated somatic embryos (ESEs) germinated successfully on 0.7% agar medium containing 3% sucrose concentration in NN basal medium (half strength of major and minor salts) with 1 mg·l-1 of gibbrellic acid. Percentage germination and plantlet development for ESEs was higher than that of non encapsulated embryos (NSEs). In comparison to different hormones, gibberellic acid has a significant influence on the germination rate of ESEs after one week of dehydration was seen maximum at 9% sucrose and abscisic acid (1 mg·l-1) in half strength of major and minor salts in Nitsch and Nitsch medium resulting in extended storage up to 90 days without loss in germination potential and capability to regenerate into plantlets. Normally developed plantlets regenerated from ESEs were successfully adapted to soil to obtain a full grown plant.

Transcriptome analysis reveals effects of red and blue lightemitting diodes(LEDs)on the growth,chlorophyll fluorescence and endogenous plant hormones of potato(Solanum tuberosum L.)plantlets cultured in vitro

Red and blue light illumination has been reported to significantly affect plantlet growth.Potato is an important food and feed crop in the world and potato plantlet cultured in vitro plays an important role in potato production.However,few studies have documented the effects of red and blue light on the growth of potato plantlets revealed at the transcriptome level.The objective of this study was to determine the growth and physiological responses of potato plantlets cultured in vitro under monochromatic red(RR),monochromatic blue(BB)as well as combined red and blue(RB)LEDs using the RNA-Seq technique.In total,3150 and 814 differentially expressed genes(DEGs)were detected in potato plantlets under RR and BB,respectively,compared to RB(used as control).Compared to the control,the DEGs enriched in"photosynthesis"and"photosynthesis-antenna proteins"metabolic pathways were up-regulated and down-regulated by BB and RR,respectively,which might be responsible for the increases and decreases of maximum quantum yield(F_(v)/F_(m)),photochemical quantum yield(φ_(PSII)),photochemical quenching(q_(P))and electron transfer rate(ETR)in BB and RR,respectively.Potato plantlets exhibited dwarfed stems and extended leaves under BB,whereas elongated stems and small leaves were induced under RR.These dramatically altered plantlet phenotypes were associated with variable levels of endogenous plant hormones gibberellin(GAs),indoleacetic acid(IAA)and cytokinins(CKs),as assessed in stems and leaves of potato plantlets.In addition,monochromatic red and blue LEDs trigged the opposite expression profiles of DEGs identified in the"plant hormone signal transduction"metabolic pathway,which were closely related to the endogenous plant hormone levels in potato plantlets.Our results provide insights into the responses of potato plantlets cultured in vitro to red and blue LEDs at the transcriptomic level and may contribute to improvements in the micro-propagation of potato plantlets cultured in vitro from the light spectrum aspect.

Effects of Lanthanum and Europium on Rooting of Plantlet Eriobotrya Japonica Lindl. in vitro

The effect of La 3+ and Eu 3+ on the rooting of Eriobotrya japonica Lindl. plantlet in vitro was studied with adding La 3+ and Eu 3+ to the rooted medium. The rooting rate, the number of root and the length of root were studied after transplanting 20 d. The activity of peroxidase, nitrate reductase and fresh weight of roots were determined after transplanting 44 d. The results show that the optimum concentration range of La 3+ (1.0～3.0 μmol·L -1), Eu 3+ (2.0～3.0 μmol·L -1) in the rooted medium can increase the rooting rate and the fresh weight of roots, and promote the length of root and raise the activities of peroxidase and nitrate reductase significantly. La 3+ has more effect in improving the rooting rate, root length and the activities of peroxidase and less effect in promoting root fresh weight and the activities of nitrate reductase than Eu 3+.

REGULATION OF PHYTOHORMONS ON THE REGENERATION OF SHOOT-TIPS OF ANTIRRHINUM MAJUS INTO PLANTLET IN VITRO

Tissue culture of Anlirrhium majus was experimented by using the tip of shoots asthe explants,and comparing the effects of NAA,IAA and 2,4—D on the formation of adventitiousbuds.The results indicated that the effect of NAA was the best formation of adventitiousbuds,5.0,7.0,9.0,11.0,13.0 mg/L BA combinated with 0.1,0.5,1.0,1.5,2.0 mg/L NAArespectively.The results show that combinations of the concentrations of BA 7.0—9.0 mg/L andNAA 0.1--0.5 mg/L were advantageous to the regeneration of buds and NAA was very importantfor callus growth.

Effect of Lanthanum on Deferring Root Ageing of Peach Plantlet in Vitro

Poor adventitious root formation is a major obstacle in micropropagation and in conventional propagation. The effects of LaCl3 on root growth and deferring the root ageing of peach plantlet in vitro were investigated. The results show that the optimum concentration of LaCl3 （2.5 μmol·L -1） in the rooted medium can significantly increase the rooting rate, the root length and the fresh weight, promote the activities of superoxide dismutase （SOD）; catalase （CAT） and peroxidase （POD） and decrease the production rate of O2 ·-, the malondialdehyde （MDA） contents and the plasma membrane permeability. There are important theory meaning and practical value in applying LaCl3 in the rooting medium to raise the rate of rooting and transplant for stone fruit tree.

Effects of Sion Cold Resistance of Cymbidium Plantletsin vitro

To determine the medium with the concentration of Na2SiO3 , the effect of different concentrations of Na2SiO3 on cold resistance of Cymbidium plantlets in vitro was studied. The results showed that Si enhanced osmotic regulation ability of Cymbidium evidently on cold resistance with increasing contents of free Pro, soluble sugar, soluble protein and decreasing contents of MDA. Compared with CK, osmotic regulation substances are accumulated rapidly. Free Pro, soluble sugar, soluble protein, MDA could be adopted as reference indices to evaluate capability of frost resistance of Cymbidium. By experiment we stated Na2SiO3 (3mmol/L) could relieve low temperature injury of Cymbidium plantlets in vitro. Mitigative effect was reduced with increasing concentration of Na2SiO3 .

Generation of Virus Free Potato Plantlets through Meristem Culture and Their Field Evaluation

Different aspects of micropropagation through meristem culture for the production of virus indexed source plants, in vitro tuberization and field evaluation of the in vitro regenerated plants were studied on four commercial cultivars of potato (Solanum tuberosum L.) viz., Diamant, Cardinal, Shilbilati and Lalpakri. The investigation was conducted at Rajshahi, Bangladesh from December 2010 to March 2012 to produce virus-free potato plantlets through meristem culture, shoot multiplications with root induction as well as their acclimatization and evaluation of morphological characters and tuber yield under field condition. Shoot tips of 25 - 30 day old field-grown plants of above mentioned four cultivars were used for meristem isolation. After isolation, meristems of these varieties of potato were cultured on “M” shaped filter paper bridge in Murashige and Skoog (MS) liquid medium. Four different treatments of media formulations viz. 0.1 mg/L KIN + 0.1 mg/L GA3, 0.1 mg/L KIN + 0.5 mg/L GA3, 0.5 mg/L KIN + 0.1 mg/L GA3 and 0.5 mg/L KIN + 0.5 mg/L GA3 were used as plant growth regulators. From these formulations MS + 0.1 mg/L KIN + 0.5 mg/L GA3 was found to be the best for the primary establishment of meristem culture. The primarily established meristems were subcultured on to MS semisolid basal medium supplemented with four different treatment combinations of hormones viz. 0.5 mg/L BA + 1.0 mg/L IBA;0.1 mg/L KIN + 0.1 mg/L GA3;0.5 mg/L BA + 0.5 mg/L GA3 and 0.5 mg/L KIN + 0.5 mg/L GA3 were used to identify the suitable media compositions for shoot proliferation. Results showed that out of these four media treatments the formulation 0.5 mg/L BA + 0.5 mg/L GA3 was found to be the best suitable for shoot generation. Among the four cultivars of potato higher frequency of shoot proliferation (number of shoots/explant and longest shoot length) was observed in Diamant, though the highest shoot formation (76%) was recorded in Cardinal. Virus free in vitro grown potato plantlets were obtained through DAS-ELISA test and used substantially for micro-propagation. After gradual acclimatization of rooted plantlets of four potato cultivars, they were transferred into the field for cultivation and established successfully. It was observed from the field study of in vitro meristem-derived plantlets that there were no virus-affected plants. The virus-free exotic varieties were much superior in all vegetative attributes and yield compared to those of indigenous varieties with producing potato plants of normal height. In contrast, the indigenous varieties took a longer time to tuber initiation and maturity, lower plant height and number of leaves per plant, a higher number of tubers but a lower amount of tuber weight per plant, and poorer tuber grade than the exotic varieties. However, the variety Cardinal exposed the best performances in the context of survival percentage of plantlets (90%), days to tuber initiation (DTI), a number of leaves per plant (NL), tuber weight per plant (343.40%) and the percentage of rich tuber grade.

Effects of Non-Conventional Substrate and NPK Fertilizer on Cassava Plantlets Acclimatization for Plant Material Production

The adaptation of cassava plantlets to natural conditions remains the main obstacle in the process of free virus plantlets production. In this study, cassava plantlets acclimatization was assessed by pre-acclimatization, height of acclimated plantlets, substrate, and nutrient watering solution. The results revealed that pre-acclimatization gave a high survival rate (61.9%) whereas the direct tunnel acclimatization was conducted to low rate (27.31%) of plantlets. Furthermore, the height of plantlets influenced significantly (p < 0.05) their survival rate with best survival rate observed on plantlet height from 1 cm to 5 cm. There was no significant difference between the substrates used and watering solutions. However, the Sawdust substrate gave a high survival rate (46.67%) and better phylogenesis (1.8) in the variety RB 89509. Likewise, plantlets treated with NPK nutrient watering solutions (20-10-10) had a better survival rate (48.57%), better phylogenesis (2.125) with the variety RB 89509 and better growth in height (1.18 cm) with the variety BF92/0267. These findings constitute a database for the establishment of the technical itinerary for the acclimatization of cassava plantlets.

Influence of Watering Solution and Phenotype on the Growth of <i>in Vitro</i>Propagated Pineapple (Smooth Cayenne Cultivar) Plantlets during Acclimatization

Pineapple is the first fruit crop cultivated in south Benin that greatly contributes for food and nutritional security and farmers’ income. But the lack of homogenous planting material constitutes the major constraint for improving pineapple yield. In vitro micropropagation is now used in the production of homogenous and free disease planting materials of pineapple. However, the acclimatization to natural condition of pineapple plantlets is an important step in planting material production of this crop. Here, we determined the intrinsic and extrinsic factors which influence the behavior of plantlets during the acclimatization process. For this purpose, plantlets from different categories were selected, trimmed and planted on a horticultural substrate made up of potting soil, white sawdust and compost previously sterilized. The plantlets were then incubated in under acclimatization greenhouse with average temperature of 29˚C and 70.2% of humidity. A batch of plantlets was subjected to two different watering solutions: Shive and Robbins solution and NPK 14-6-5 foliar fertilizer. The results obtained initially showed high rate (100%) of survival and growth of the plantlets watered with Shive and Robbins solution against 50% of the plantlets watered with the foliar fertilizer solution. In addition, the plantlets with spread pores exhibiting the characteristics of which the number of leaves varies between 9 and 11, the weight between 1.2 and 1.5 g, the size of 4.5 to 5.5 cm, and a good junction between the aerial part and the root system were those which were successfully grown in acclimatization phase under greenhouse, unlike plantlets with erected pores having lower success rate. This study goes a long way in providing good procedures of acclimatization of homogenous and free disease planting material of pineapple to the famers.

Study on Efficient Multiplication of Blueberry(Vaccinium corymbosum)Plantlets in Swinging type Temporary Immersion Bioreactors System(S-TIBs)

The swinging type temporary immersion bioreactors system( S-TTBs) is a kind of new and advanced method of tissue culture. The efficient multiplication technology of blueberry( Vaccinium corymbosum) plantlets in S-TTBs was systematically studied through L_9( 3~4) orthogonal experiment with three factors( culture medium volume,swing angle,and inoculation density) and completely randomized experiment with two factors( swing frequency and immersion time).The results showed that the optimal culture parameters were set as follows: the culture medium volume of 250 ml/bottle,the swing angle at 45°,and the inoculation density of 60 plantlets/bottle. The optimal swing frequency was 1 time/6 h,and the immersion time was set as 60 s.

Effects of mechanical vibration on root development of Actinidia chinensis plantlet

The root development of Actinidia chinensis plantlets was studied in exposure to environmental stress of mechanical vibration at respectively 1 Hz, 2 Hz, 3 Hz, 4 Hz and 5 Hz. The plantlets exposed to vibration stimuli at all those frequencies have a larger total number and a larger total length of roots and a smaller permeability of root plasma-membrane, compared with those cultivated in an environment without vibration stress. Vibration at respectively 1 Hz, 2 Hz, 3 Hz and 4 Hz enhances root activity and the 3 Hz vibration is the most favorable. There is an obvious negative correlation between root activity and permeability of root plasma-membrane. The effects may be explained by the likelihood that mechanical vibration at an appropriate frequency facilitates roots’ absorbing water and minerals which are indispensable to inducing and synthesizing in roots some active substances favorable to growth. Nevertheless, overstress damages the integrity of root plasm-membrane, increases the permeability, and results in the disability of protecting root cells.

Construction and Characterization of cDNA Library from Water-Stressed Plantlets Regenerated in vitro of Populus hopeiensis

In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by LD-PCR with the SMART cDNA Library Construction Kit, was n vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68×10i6 and 1.69×109 pfu·mL–1 respectively. The combination ratio reached 98.8% and the average size of inserts was about 800 bp. In addition, the percentage of inserted fragments (> 400 bp) was approximately 90%. The results indicate that a cDNA library has been successfully constructed.